Project description:Germline-competent embryonic stem cells (ESCs) have been derived from mice and rats, but not from other species. Here, we report the development of conditions for the derivation of ESCs from chickens and several other avian species. Chicken ESCs express core pluripotency markers and can efficiently differentiate into cells of all three germ layers. Furthermore, chicken ESCs can form high rates of chimerism when injected into chicken embryos both ex ovo and in ovo. More importantly, chicken ESCs can give rise to germ cells both in vitro and in ovo, indicating that chicken ESCs are germline competent. Interspecies chimeras can also be generated by injecting other avian ESCs into chicken embryos. Establishment of germline competent avian ESCs opens up a new avenue for producing genetically engineered avian species.
Project description:Chimeric and germline-competent embryonic stem cells (ESCs) have been derived from mice and rats, but not from other species. Here, we report the development of conditions for the derivation of ESCs from chickens and several other avian species. Chicken ESCs express core pluripotency markers and can efficiently differentiate into cells of all three germ layers. Furthermore, chicken ESCs can form high rates of chimerism when injected into chicken embryos both ex ovo and in ovo. More importantly, chicken ESCs can give rise to germ cells both in vitro and in ovo, indicating that chicken ESCs are germline competent. Interspecies chimeras can also be generated by injecting other avian ESCs into chicken embryos. Establishment of germline competent avian ESCs opens up a new avenue for producing genetically engineered avian species.
Project description:Hypoblast stem cells were derived from rat embryonic stem cells (rESCs) by culturing with LIF, EGF, PDGF, and 2% fetal bovine serum. mRNA expression profiles of the cells during derivation at different time points were determined with Affymetrix microarrays. Global transcriptome of rESC-derived hypoblast stem cells (cHypoSCs) was similar to that of rat embryo-derived hypoblast stem cells and rat multipotent adult progenitor cells. In addition, the transcriptome of these cells were compared with the previously published datasets including transcriptome of mouse embryonic stem cells, extraembryonic endoderm cells, single cells of mouse embryos (E3.25, E3.5, and E4.5). The comparison revealed that cHypoSCs have the transcriptome similar to that of extraembryonic endoderm cells and E3.5 primitive endoderm cells.